Study on the Effects of Probiotics on the Growth ofCyprinus carpiodand Water Quality

Wanqing GUO,Lidan BAI,Hai WANG,Xiaowei LI,Wei SHI,Shuo SUN

1.Changchun Aquatic Products Quality Safety Testing Center,Changchun 130033,China;

2.Changchun Modern Agricultural Park,Changchun 130118,China;

3.Yushu Aquatic Product Technology Promotion Department,Yushu 130400,China

Study on the Effects of Probiotics on the Growth ofCyprinus carpiodand Water Quality

Wanqing GUO1*,Lidan BAI1,Hai WANG1,Xiaowei LI2,Wei SHI2,Shuo SUN3

1.Changchun Aquatic Products Quality Safety Testing Center,Changchun 130033,China;

2.Changchun Modern Agricultural Park,Changchun 130118,China;

3.Yushu Aquatic Product Technology Promotion Department,Yushu 130400,China

[Objective]The aim was to discuss the effects of probiotics on the growth ofCyprinus carpiodand water quality.[Method]TakingC．carpiodas the research object,probiotics were supplemented in the fodder and water to study their effects on the growth ofC．carpiodand water quality.[Result]Probiotics had promoting effects on the growth ofC．carpiodand its optimum dosage was 6%．pH,ammonia nitrogen content and nitrite content in water body in experimental groups were all lower than those in control group.[Conclusion]Compound probiotics had a broad application foreground in the aquatic breeding industry．

Cyprinus carpiod;Probiotics;Growth;Ammonia nitrogen;Nitrite

P robiotics are a normal flora and metabolite improving the health level of the host,adjusting microecological imbalance and maintaining microecological balance, they are also a generic name[1]of microbial ecological agents promoting the growth of normal flora selectively in the guidance of ecological theory.In addition,they are a green additive as well,they show significant effects on promoting the growth and development of animal,purifying water quality, enhancing immunity,perfecting the palatability of fodder and controlling medicinal residue,etc.[2],what’s more, they can solve the problems of food safety,environmental protection and human health,meeting the needs of sustainable development,thus it is an effective approach to coordinate the relationship between human and nature,and promote the development of aquaculture industry.

Bacillusis a single fungus belonging to probiotics.As a kind of aerobic bacteria,it can produce spores with the features of acid-base resistance, high-temperature resistance and cholate resistance under certain conditions,thus it can tolerate the production and processing of granule.This kind of microbial inoculum often has the following functions:first,it can generate lots of nutrient substances, such as amylase,protease,lipase, amino acid,vitamin and somatomedin,etc.,these can promote the digestion and absorption of nutrient substances; second,microbial oxygen consumption,which can keep an anaerobic environment for gastrointestinal tract,it is often mixed and used with anaerobic probiotics like lactic acid bacteria,etc.; third,biological antagonism,which will inhibit the growth of pathogenic bacteria,and adjust ecological balance of intestinal tract;fourth,it can accelerate lymphopoiesis and immune globulin secretion,thereby improving the immunologic function of animal organism.There are many bacilli suitable for the production of microorganism feed supplement,such asBacillus subtilis,Bacillus cereus,Bacillus licheniformisandBacillus toyoi,etc.[3-6].

In the middle 1970s abraod,probiotics have already been used in livestock feed.Since the 1980s at home, livestock probiotics have been gradually used,moreover,there have been few reports on the use of probiotics in fish culture[7].Previous studies have already shown that probiotics had good promoting effects on the growth of fish[8].Cyprinus carpiod,as a rare large-scale freshwater ornamental fish,has the good reputation of“the King of Aquarium Fish”,of course it has very high ornamental value and economic value.In this paper,takingC．carpiodas the research object,probiotics(compound bacilli)were splashed and fed in aquarium fish ponds to further study their effects on the growth of aquarium fish and water quality in the breeding process.

Materials and Methods

Experimental materials

Experimental fishAllC．carpiodfor the experiment were healthy,the weight and the length of each were (87.23±0.43)g and(13.27±0.54)cm, respectively,both eligible male and eligible female can be used,10 mg/L potassium permanganate solution must be used for rinsing and sterilizing before experiment.

Experimental instrumentsThe instruments for the experiment were:4 oblong transparent glass fishbowls, the water should be stood in the fish tank for one day,aerating and oxygenating for 24 h;dividing rule;electronic scales;dissecting tray;scrape; acidimeter;quick test box of sulfuretted hydrogen;rapid detectionkit of water quality;quick test box of COD;rapid detection kit of hardness of water.

ProbioticsThe probiotics for the experiment were compound bacillus powder,its main components are Bacillus subtilis,Bacillus natto,Bacillus licheniformis and Bacillus cereus, as well as some adjuvants like biological enzyme,vitamin and microelement,total number of beneficial bacteria in vivo was≥1×108per g.

Basal feedBasal feed was a special compound feed for C．carpiod,its principal raw materials were refined imported fish oil,imported white fish powder,imported steam brown fishmeal (TVBN＜110),senior shrimp powder, high protein flour,mineral substance, multivitamin,compound feeding attractant,immunopotentiator and probiotics and so on.The nutrient level of basal feed was in Table 1.

According to total number and specification of C．carpiod culture in the tank,daily feeding amount was 3% of fish weight,and it should be fed twice each day at 8:30 a.m.and 1:30 p.m.,the culture cycle was 30 d.

Experimental methods

Experimental designThe comparison method between experimental tanks and control tank was adopted. This two kinds of tanks had the fish with the same size,specification, quantity,water volume,and feeding and management methods.Probiotics were used in experimental tanks and not in control tank.

(1)No.1,2 and 3 were experimental tanks.①External use:Soaking probiotics in aquaculture water for 5 h, then they were evenly splashed into the tanks every five days,2 g/m3probiotics were used for each time.②To be taken orally:20,60 and 100 g probiotics were respectively added into fodder per kg,feeding according to conventional method.Probiotics can not be stored and used with bactericide,disinfector and chemical medicine,etc.

(2)No.4 was control tank.Don’t use probiotics,feeding and managing normally.①Water samples from experimental tanks and control tank were both collected.Before splashing probiotics each time,water samples of experimental tanks and control tank should be collected every 5 d, moveover,each sampling time and site were fixed.Then dissolved oxygen,sulfuretted hydrogen,pH,ammonia nitrogen,nitrite,nitrate,potassium permanganate index,total hardness and total alkalinity should be measured respectively.②5 fish were sampled randomly from No.1,2,3,4 tanks respectively,then the overall length, body length,height and weight of these fish were measured and recorded respectively.③The measured data and management situation of eack tank were recorded.④30 d after the experiment,the overall length,body length,height and weight of these fish were measured again.

Data processing

EXCEL software was used to calculate average value and standard deviation(x±SD)of all data,and SPSS17.0 statistical software was used to carry out one-way analysis of variance on experimental data.If there were significant differences, LSD multiple comparisons were carried out,the significance level was P＜0.05.Growth rate and weight gain rate were calculated according to the following formulae:

Growth rate=(Average final body length-Average initial body length)/Average initial body length×100%(1)

Weight gain rate=(Average final weight-Average initial weight)/Average initial weight×100%(2)

Table 1The nutrient level of basal feed %

Results and Analyses

Effects of probiotics on the growth ofC．carpiod

The experimental duration was 30 d(from March 1 to March 31, 2013),the experimental results were in Fig.1,Fig.2 and Table 2.

Seen from Fig.1,Fig.2 and Table 1,it can be inferred that probiotics of different concentrations had extremely significant promoting effects on the growth of C．carpiod(P＜0.01),there was no significant difference between 2%and 10%probiotics groups,and other groups had extremely significant differences(P＜0.01),in which weight gain rate of C．carpiod in 6%probiotics group was 51.41%and the efffect was the most obvious;weight gain rates of 2%probiotics group and 10% probiotics group were 34.20%and 33.93%,respectively,the two groups had similar promoting effects on the growth of C．carpiod.Thus it can be seen that probiotics had promoting effects on the growth of C．carpiod, moreover,the effects followed the following sequence:6%probiotics group＞2%probiotics group＞10% probiotics group.

Effects of probiotics on water quality

pHAs shown in Fig.3,the change range of pH of each experiment group was small,and p H showed declining trend,indicating that probiotics can fully decompose organic matter and maintain the stablility of pH.pH of control group was among 6.48-7.43, the fluctuation was large,it decreased at the initial stage and raised in the middle and later periods.

NitriteAs indicated in Fig.4,nitrate content of control group was higher than that of experimental groups all the time,it had a slow growth trend at the initial stage,while had a quick growth trend at the later stage.For experimental groups,it showed a slow growth trend at the initial stage,and had a large rising range at the later stage.These showed that probiotics had limited ability to decompose nitrite.

Table 2Effects of probiotics on the growth ofC．carpiod

Ammonia nitrogen contentAs inferred in Fig.5,for control group, ammonia nitrogen content had a slow growth trend at the initial stage,and showed a trend of first increasing rapidly and then decreasing in the middle and later periods,and the variation range was large.For experimental groups,it had a slow growth trend,moreover,the variation range was small.These inferred that probiotics can effectively lower ammonia nitrogen content in water.

Discussions

Effects of probiotics on the growth ofC．carpiod

Probiotics can fix in the intestinal tract and saturate the adsorption site of intestinal mucosa,causing pathogenic bacteria can not fix in intestinal mucosa,thereby maintaining ecological balance.Its mechanism of stimulating the growth was generating vitamin and amino acid.Probiotics contained lots of nutrient substances, with the fixing,propagation and metabolism of them in the digestive tract of animals,they helped somatomedin,protease,lipase,amylase and cellulase[9-10]participate in some important metabolic reactions of organism,thereby promoting the absorption of nutrient substances like vitamin,etc.,cooperating with the host to increase the digestion and absorption of fodder,moreover,thallithemselves of beneficial bacterium contained lots of nutrient substances, which can supplement the nutritions for the growth and development of aquatic livestock,thus promoting the growth ofC．carpiod[11].

The experiment showed that probiotics of different concentrations had extremely significant promoting effects on the growth ofC．carpiod(P＜0.01),in which weight gain rates ofC．carpiodin 6%,2%and 10%probiotics groups were 51.41%,34.20% and 33.93%,respectively,obviously, the effect of 6%probiotics group was the most obvious,the other two groups had similar promoting effects on the growth ofC．carpiod.Thus it can be seen that probiotics had promoting effects on the growth ofC．carpiod, moreover,the effects followed the following sequence:6%probiotics group＞2%probiotics group＞10%probiotics group.The outstanding effect of 6%probiotics group may relate to beneficial bacterium which can quickly enter into and fix in the intestinal tract ofC．carpiodunder 6%concentration of probiotics,thereby improving the digestibility ofC．carpiod[12].

Effects of probiotics on water quality

Probiotics added into fodder can be directly fixed in the intestinal tract of animals,and probiotics which had not been fixed and digested would be passed out of the body in the faeces[18]. In improving water quality,the additive amount of probiotics in water should be higher than that in fodder theoretically,which was beneficial to the improvement of water quality. Compound probiotics can not only alleviate the pollution of breeding environment,achieving the effect of preventing diseases and healthy breeding,but also because of the nutrition of thalli themselves,it can supplement the nutritions for the growth and development of aquatic livestock.In sum,compound probiotics had a broad application foreground in the aquatic breeding industry[19].

[1]LIAN XH(廉新慧),ZHANG GN(张高娜), LIU FZ(刘凤芝),et al.Application ofBacillus subtilisin animal production(枯草芽胞杆菌在动物生产中的应用)[J]. Animal Husbandry and Feed Science (畜牧与饲料科学),2012,33(10):23-24.

[2]ZHANG RS(张瑞霜).Inheriting the concrete matters,cooperating and innovating,new direction of probiotics study and application(传承务实合作创新聚焦微生态制剂研究应用新方向——第二届饲料微生态制剂应用技术研讨会暨微生态制剂大会纪实)[C].//ZHANG RS(张瑞霜).Feed and Husbandry(饲料与畜牧).Beijing:Beijing Feed Industry Association(北京:北京市饲料工业协会),2012:46.

[3]DOU MX(窦茂鑫),WU T(吴涛).Study on the development status and application of feeding probiotics(饲用微生态制剂的发展现状与应用性研究)[J].Feed Research(饲料研究),2013,(1):13-17.

[4]CAO AQ(曹爱青).Study on the application of animal probiotics in animal husbandry(动物微生态制剂在畜牧业中的应用研究)[J].Animal Health (动物保健),2012(20):37-39.

[5]WANG Y(王芸),ZHENG ZL(郑宗林). The application research progresses of microbial ecological preparation in aquaculture(微生态制剂在水产养殖中的应用研究进展)[J].Feed and Husbandry(饲料与畜牧),2013,(2):18-21.

[6]KOZASA M.Toyocerin(Bacillus toyoi) as growth promotor for ani mal feeding [J].Microbiologie Aliments Nutrition, 1986,4(1):121-135.

[7]XIANG DC(向德超),SONG WP(宋维平),LIN H(林红),et al.Effects of adding probiotics into fish bait to the growth of grass carp(饵料中添加微生态制剂对草鱼生长的影响)[J].Journal of Southwest Agricultural University:Social Science Edition(四川畜牧兽医学报),1995(2): 12-16.

[8]ZHOU X,TIAN Z,WANG Y,et al.Effect of treatment with probiotics as water additives on tilapia(Oreochromis niloticus)growth performance and immune response[J].Fish Physiology Biochemistry,2010,36(3):501-509.

[9]YANG HP(杨蕙萍),TONG SY(童圣英), WANG ZC(王子臣).Overview of digestive enzyme research of aquatic livestock at home and abroad(国内外关于水产动物消化酶研究的概况)[J].Journal of Dalian Fisheries University(大连水产学院学报),1998,13(3):64-71.

[10]DING X(丁贤),LI ZJ(李卓佳).Effects of bacillus on the growth ofLitopenaeus vannameiand the activity of digestive enzyme(芽孢杆菌对凡纳滨对虾生长和消化酶活性的影响)[J].Journal of Fishery Sciences of China(中国水产科学),2004,11(6):6-11.

[11]ZHU XZ(朱学芝),ZHENG SX(郑石轩),PAN QJ(潘庆军).Effect of probiotics on growth performance and water quality forLitopenaeus vannamei(微生态制剂对凡纳滨对虾生长及水质的影响)[J].Acta Scientiarum Naturalium Universitatis Sunyatseni(中山大学学报),2008(z1):58-62.

[12]WU P(武鹏),ZHAO DQ(赵大千),CAI HH(蔡欢欢),et al.Effects of three probiotics on water quality and growth in sea cucumberApostichopus japonicus(3种微生态制剂对水质及刺参幼参生长的影响)[J].Journal of Dalian Fisheries University(大连海洋大学学报), 2013,28(1):21-26.

[13]WU BC(吴保承),SHEN GQ(沈国强), YANG CX(杨春霞),et al.The application status and prospects about the probiotics used in water purification(微生态制剂在水质净化中的应用现状及展望)[J].Environmental Science and Technology(环境科学与技术),2010, 33(S2):408-410.

[14]WANG DC(王笃彩),YAN BL(闫斌伦), LI SH(李士虎),et al.Comparison of influence of three micro-ecological pharmaceutics on cultured water quality(3种微生态制剂对养殖水体水质影响的比较研究)[J].Journal of Hydroecology(水生态学杂志),2011,32(1): 66-70.

[15]GAI JJ(盖建军),JIAO XM(矫新明), CHEN HG(陈焕根).Effects of four probiotics on water quality of fishpond(4种微生态制剂对养殖水质的影响)[J]. Modern Agricultural Science and Technology(现代农业科技),2013(10): 255-256.

[16]LIU TG(刘铁钢),ZHAO W(赵文),LIU G(刘钢),et al.The effects of five probiotics on the ecological security of sea cucumber Apostichopus japonicus juveniles(5种微生态制剂对刺参幼参的生态安全性)[J].Journal of Dalian Fisheries University(大连海洋大学学报),2012,27(2):129-136.

[17]XU Q(徐琴),LI J(李健),LIU Q(刘淇),et al.The effects of four kinds of microbial ecological agent on water quality of Fenneropenaeus chinensis larvae (4种微生态制剂对对虾育苗水体主要水质指标的影响)[J].Marine Sciences (海洋科学),2009,33(3):10-15.

[18]HU Y(胡毅).Optimum feed formula selection and some feed additive application on juvenile litopenaeus vannamei(凡纳滨对虾饲料配方优化及几种饲料添加剂的应用)[D].Qingdao:Ocean University of China(青岛:中国海洋大学),2007:133-139.

[19]CHEN QH(陈秋红),SHI DL(施大林), LV HM(吕惠敏),et al.Cleansing effects of compound microecological preparation to breed bodies of water (复合微生态制剂对水产养殖水体净化作用的研究)[J].Biotechnology(生物技术),2004,14(4):63-64.

Responsible editor:Nanling WANG

Responsible proofreader:Xiaoyan WU

Supported by the Project of Quality Safety of Agricultural Products Loaned from World Bank of Jilin Province.

About the Author:Wanqing GUO,male,was born in May 1963,graduated from Dalian Ocean University,Research Fellow,mainly interested in the analysis of new fish species and water quality of fish culture.

*Corresponding author.E-mail:Haowanghai@163.com

Received:March 20,2015 Accepted:May 15,2015